Transport of the sulfated, amidated bile acid, sulfolithocholyltaurine, into rat hepatocytes is mediated by Oatp1 and Oatp2

Ling Jie Meng, Pijun Wang, Allan W. Wolkoff, Richard B. Kim, Rommel G. Tirona, Alan F. Hofmann, K. Sandy Pang

Research output: Contribution to journalArticle

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Abstract

The uptake of the sulfated bile acid sulfolithocholyltaurine (SLCT) was investigated in isolated rat hepatocytes and in HeLa cells transfected with complementary DNAs (cDNAs) of organic anion transporting polypeptides (Oatps) 1 and 2 cloned from rat liver. In hepatocytes, transport of SLCT was greatly reduced by bromosulfophthalein (BSP), estrone sulfate, the precursor bile acids cholyltaurine and lithocholyltaurine, and 4,4′-diisothiocyanostilbene-2-2′-disulfonic acid (DIDS). However, SLCT transport was insensitive to 4-methylumbelliferyl sulfate, harmol sulfate, digoxin, fexofenadine, and lack of sodium ion. Because the estimation of kinetic constants was enhanced with use of inhibitors, BSP (1-50 μmol/L) was added to isolated rat hepatocytes to assess the various transport components for SLCT uptake. The resulting data showed a nonsaturable pathway and at least 2 pathways of different Michaelis-Menten constants (Km) (70 and 6 μmol/L) and similar maximum velocities (Vmax) (1.73 and 1.2 nmol/min/mg protein) and inhibition constants of 0.63 and 10.3 μmol/L for BSP. In expression systems, SLCT was taken up by Oatp1 and Oatp2 expressed in HeLa cells with similar Km values (12.6 ± 6.2 and 14.6 ± 1.9 μmol/L). These Km values were comparable to that observed for the high-affinity pathway in rat hepatocytes. In conclusion, the results suggest that transport of SLCT into rat liver is mediated in part by Oatp1 and Oatp2, high-affinity pathways, a lower-affinity pathway of unknown origin, and a nonsaturable pathway that is compatible with a transport system of high Km and/or passive diffusion.

Original languageEnglish (US)
Pages (from-to)1031-1040
Number of pages10
JournalHepatology
Volume35
Issue number5
DOIs
StatePublished - 2002

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Bile Acids and Salts
Hepatocytes
Sulfobromophthalein
fexofenadine
HeLa Cells
Taurolithocholic Acid
Taurocholic Acid
Liver
Digoxin
Anions
Complementary DNA
Sodium
Ions
Peptides
Acids
Proteins

ASJC Scopus subject areas

  • Hepatology

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Transport of the sulfated, amidated bile acid, sulfolithocholyltaurine, into rat hepatocytes is mediated by Oatp1 and Oatp2. / Meng, Ling Jie; Wang, Pijun; Wolkoff, Allan W.; Kim, Richard B.; Tirona, Rommel G.; Hofmann, Alan F.; Pang, K. Sandy.

In: Hepatology, Vol. 35, No. 5, 2002, p. 1031-1040.

Research output: Contribution to journalArticle

Meng, Ling Jie ; Wang, Pijun ; Wolkoff, Allan W. ; Kim, Richard B. ; Tirona, Rommel G. ; Hofmann, Alan F. ; Pang, K. Sandy. / Transport of the sulfated, amidated bile acid, sulfolithocholyltaurine, into rat hepatocytes is mediated by Oatp1 and Oatp2. In: Hepatology. 2002 ; Vol. 35, No. 5. pp. 1031-1040.
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abstract = "The uptake of the sulfated bile acid sulfolithocholyltaurine (SLCT) was investigated in isolated rat hepatocytes and in HeLa cells transfected with complementary DNAs (cDNAs) of organic anion transporting polypeptides (Oatps) 1 and 2 cloned from rat liver. In hepatocytes, transport of SLCT was greatly reduced by bromosulfophthalein (BSP), estrone sulfate, the precursor bile acids cholyltaurine and lithocholyltaurine, and 4,4′-diisothiocyanostilbene-2-2′-disulfonic acid (DIDS). However, SLCT transport was insensitive to 4-methylumbelliferyl sulfate, harmol sulfate, digoxin, fexofenadine, and lack of sodium ion. Because the estimation of kinetic constants was enhanced with use of inhibitors, BSP (1-50 μmol/L) was added to isolated rat hepatocytes to assess the various transport components for SLCT uptake. The resulting data showed a nonsaturable pathway and at least 2 pathways of different Michaelis-Menten constants (Km) (70 and 6 μmol/L) and similar maximum velocities (Vmax) (1.73 and 1.2 nmol/min/mg protein) and inhibition constants of 0.63 and 10.3 μmol/L for BSP. In expression systems, SLCT was taken up by Oatp1 and Oatp2 expressed in HeLa cells with similar Km values (12.6 ± 6.2 and 14.6 ± 1.9 μmol/L). These Km values were comparable to that observed for the high-affinity pathway in rat hepatocytes. In conclusion, the results suggest that transport of SLCT into rat liver is mediated in part by Oatp1 and Oatp2, high-affinity pathways, a lower-affinity pathway of unknown origin, and a nonsaturable pathway that is compatible with a transport system of high Km and/or passive diffusion.",
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T1 - Transport of the sulfated, amidated bile acid, sulfolithocholyltaurine, into rat hepatocytes is mediated by Oatp1 and Oatp2

AU - Meng, Ling Jie

AU - Wang, Pijun

AU - Wolkoff, Allan W.

AU - Kim, Richard B.

AU - Tirona, Rommel G.

AU - Hofmann, Alan F.

AU - Pang, K. Sandy

PY - 2002

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AB - The uptake of the sulfated bile acid sulfolithocholyltaurine (SLCT) was investigated in isolated rat hepatocytes and in HeLa cells transfected with complementary DNAs (cDNAs) of organic anion transporting polypeptides (Oatps) 1 and 2 cloned from rat liver. In hepatocytes, transport of SLCT was greatly reduced by bromosulfophthalein (BSP), estrone sulfate, the precursor bile acids cholyltaurine and lithocholyltaurine, and 4,4′-diisothiocyanostilbene-2-2′-disulfonic acid (DIDS). However, SLCT transport was insensitive to 4-methylumbelliferyl sulfate, harmol sulfate, digoxin, fexofenadine, and lack of sodium ion. Because the estimation of kinetic constants was enhanced with use of inhibitors, BSP (1-50 μmol/L) was added to isolated rat hepatocytes to assess the various transport components for SLCT uptake. The resulting data showed a nonsaturable pathway and at least 2 pathways of different Michaelis-Menten constants (Km) (70 and 6 μmol/L) and similar maximum velocities (Vmax) (1.73 and 1.2 nmol/min/mg protein) and inhibition constants of 0.63 and 10.3 μmol/L for BSP. In expression systems, SLCT was taken up by Oatp1 and Oatp2 expressed in HeLa cells with similar Km values (12.6 ± 6.2 and 14.6 ± 1.9 μmol/L). These Km values were comparable to that observed for the high-affinity pathway in rat hepatocytes. In conclusion, the results suggest that transport of SLCT into rat liver is mediated in part by Oatp1 and Oatp2, high-affinity pathways, a lower-affinity pathway of unknown origin, and a nonsaturable pathway that is compatible with a transport system of high Km and/or passive diffusion.

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